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Hormone PDB id
6ins
Jmol
Contents
Protein chains
50 a.a. *
Metals
_ZN ×2
Waters ×179
* Residue conservation analysis
PDB id:
6ins
Name: Hormone
Title: X-ray analysis of the single chain b29-a1 peptide-linked ins molecule. A completely inactive analogue
Structure: Insulin. Chain: e, f. Engineered: yes
Source: Sus scrofa. Pig. Organism_taxid: 9823. Collection: pancreas
Resolution:
2.00Å     R-factor:   0.179    
Authors: U.Derewenda,Z.Derewenda,E.J.Dodson,G.G.Dodson,X.Bing,J.Marku
Key ref: U.Derewenda et al. (1991). X-ray analysis of the single chain B29-A1 peptide-linked insulin molecule. A completely inactive analogue. J Mol Biol, 220, 425-433. PubMed id: 1856866 DOI: 10.1016/0022-2836(91)90022-X
Date:
25-Nov-92     Release date:   31-Jan-94    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P01315  (INS_PIG) -  Insulin
Seq:
Struc: 		108 a.a.
50 a.a.
Key:    PfamA domain  Secondary structure  CATH domain

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   1 term 
  Biochemical function     hormone activity     1 term  

 

 
DOI no: 10.1016/0022-2836(91)90022-X J Mol Biol 220:425-433 (1991)
PubMed id: 1856866  
 
 
X-ray analysis of the single chain B29-A1 peptide-linked insulin molecule. A completely inactive analogue.
U.Derewenda, Z.Derewenda, E.J.Dodson, G.G.Dodson, X.Bing, J.Markussen.
 
  ABSTRACT  
 
A crystal structure of a totally inactive insulin molecule has been determined. For this insulin molecule, the first without detectable activity to be characterized, the A and B-chains are linked by a peptide bond between A1 Gly and B29 Lys. The molecule has retained all its normal self-association properties and it can also accommodate the two different conformations designated T and R, as seen in 4Zn native pig insulin crystals. The hexamers of the crosslinked insulin molecule were crystallized using the 4Zn insulin recipe of Schlichtkrull. The structure has been crystallographically refined with data extending to 2 A using restrained least-square methods. Comparison of the B29-A1 peptide crosslink insulin and the 4Zn native insulin reveals close structural similarities with the native dimer. The analysis of the structure confirms the earlier hypothesis that insulin structures in crystals are not in an active conformation and that a separation of N-terminal A-chain and C-terminal B-chain is required for interaction with the insulin receptor.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
20133841 J.Jirácek, L.Záková, E.Antolíková, C.J.Watson, J.P.Turkenburg, G.G.Dodson, and A.M.Brzozowski (2010).
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Exploring the implications of vitamin B12 conjugation to insulin on insulin receptor binding.
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19321435 B.Xu, K.Huang, Y.C.Chu, S.Q.Hu, S.Nakagawa, S.Wang, R.Y.Wang, J.Whittaker, P.G.Katsoyannis, and M.A.Weiss (2009).
Decoding the Cryptic Active Conformation of a Protein by Synthetic Photoscanning: INSULIN INSERTS A DETACHABLE ARM BETWEEN RECEPTOR DOMAINS.
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19321436 Q.X.Hua, B.Xu, K.Huang, S.Q.Hu, S.Nakagawa, W.Jia, S.Wang, J.Whittaker, P.G.Katsoyannis, and M.A.Weiss (2009).
Enhancing the Activity of a Protein by Stereospecific Unfolding: CONFORMATIONAL LIFE CYCLE OF INSULIN AND ITS EVOLUTIONARY ORIGINS.
  J Biol Chem, 284, 14586-14596.
PDB codes: 2k91 2k9r
18452211 M.Morita, S.Nakamura, and K.Shimizu (2008).
Highly accurate method for ligand-binding site prediction in unbound state (apo) protein structures.
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18332129 Q.X.Hua, S.H.Nakagawa, W.Jia, K.Huang, N.B.Phillips, S.Q.Hu, and M.A.Weiss (2008).
Design of an active ultrastable single-chain insulin analog: synthesis, structure, and therapeutic implications.
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PDB codes: 2jzq 3bxq
18071885 Z.J.Zhang, L.Wu, Z.S.Qiao, M.Q.Qiao, Y.M.Feng, and Z.Y.Guo (2008).
Contribution of the conserved A16Leu to insulin foldability.
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17280834 C.W.Ward, M.C.Lawrence, V.A.Streltsov, T.E.Adams, and N.M.McKern (2007).
The insulin and EGF receptor structures: new insights into ligand-induced receptor activation.
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Insulin structure and function.
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The A-chain of insulin contacts the insert domain of the insulin receptor. Photo-cross-linking and mutagenesis of a diabetes-related crevice.
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PDB codes: 2jum 2juu 2juv
17436342 L.Záková, D.Zyka, J.Jezek, I.Hanclová, M.Sanda, A.M.Brzozowski, and J.Jirácek (2007).
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I222 crystal form of despentapeptide (B26-B30) insulin provides new insights into the properties of monomeric insulin.
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PDB code: 2ceu
16365033 K.J.Rosengren, F.Lin, R.A.Bathgate, G.W.Tregear, N.L.Daly, J.D.Wade, and D.J.Craik (2006).
Solution structure and novel insights into the determinants of the receptor specificity of human relaxin-3.
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PDB code: 2fhw
17716170 M.Koch, F.F.Schmid, V.Zoete, and M.Meuwly (2006).
Insulin: a model system for nanomedicine?
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16894147 M.Lou, T.P.Garrett, N.M.McKern, P.A.Hoyne, V.C.Epa, J.D.Bentley, G.O.Lovrecz, L.J.Cosgrove, M.J.Frenkel, and C.W.Ward (2006).
The first three domains of the insulin receptor differ structurally from the insulin-like growth factor 1 receptor in the regions governing ligand specificity.
  Proc Natl Acad Sci U S A, 103, 12429-12434.
PDB code: 2hr7
16762918 Q.X.Hua, S.Nakagawa, S.Q.Hu, W.Jia, S.Wang, and M.A.Weiss (2006).
Toward the active conformation of insulin: stereospecific modulation of a structural switch in the B chain.
  J Biol Chem, 281, 24900-24909.
PDB codes: 2hh4 2hho
16751187 S.H.Nakagawa, Q.X.Hua, S.Q.Hu, W.Jia, S.Wang, P.G.Katsoyannis, and M.A.Weiss (2006).
Chiral mutagenesis of insulin. Contribution of the B20-B23 beta-turn to activity and stability.
  J Biol Chem, 281, 22386-22396.  
16981237 V.Zoete, and M.Meuwly (2006).
Importance of individual side chains for the stability of a protein fold: computational alanine scanning of the insulin monomer.
  J Comput Chem, 27, 1843-1857.  
16239223 K.Huang, J.Dong, N.B.Phillips, P.R.Carey, and M.A.Weiss (2005).
Proinsulin is refractory to protein fibrillation: topological protection of a precursor protein from cross-beta assembly.
  J Biol Chem, 280, 42345-42355.  
16215634 Z.Y.Guo, Z.Zhang, X.Y.Jia, Y.H.Tang, and Y.M.Feng (2005).
Mutational analysis of the absolutely conserved B8Gly: consequence on foldability and activity of insulin.
  Acta Biochim Biophys Sin (Shanghai), 37, 673-679.  
15551269 P.De Meyts (2004).
Insulin and its receptor: structure, function and evolution.
  Bioessays, 26, 1351-1362.  
15653430 Z.Y.Guo, X.Y.Jia, and Y.M.Feng (2004).
Replacement of the interchain disulfide bridge-forming amino acids A7 and B7 by glutamate impairs the structure and activity of insulin.
  Biol Chem, 385, 1171-1175.  
12930990 H.B.Olsen, M.R.Leuenberger-Fisher, W.Kadima, D.Borchardt, N.C.Kaarsholm, and M.F.Dunn (2003).
Structural signatures of the complex formed between 3-nitro-4-hydroxybenzoate and the Zn(II)-substituted R(6) insulin hexamer.
  Protein Sci, 12, 1902-1913.  
12649435 H.Yan, Z.Y.Guo, X.W.Gong, D.Xi, and Y.M.Feng (2003).
A peptide model of insulin folding intermediate with one disulfide.
  Protein Sci, 12, 768-775.  
12590147 M.Liu, J.Ramos-Castañeda, and P.Arvan (2003).
Role of the connecting peptide in insulin biosynthesis.
  J Biol Chem, 278, 14798-14805.  
12654724 Q.X.Hua, S.H.Nakagawa, J.Wilken, R.R.Ramos, W.Jia, J.Bass, and M.A.Weiss (2003).
A divergent INS protein in Caenorhabditis elegans structurally resembles human insulin and activates the human insulin receptor.
  Genes Dev, 17, 826-831.  
14573855 X.Y.Jia, Z.Y.Guo, Y.Wang, Y.Xu, S.S.Duan, and Y.M.Feng (2003).
Peptide models of four possible insulin folding intermediates with two disulfides.
  Protein Sci, 12, 2412-2419.  
14596591 Z.L.Wan, B.Xu, Y.C.Chu, P.G.Katsoyannis, and M.A.Weiss (2003).
Crystal structure of allo-Ile(A2)-insulin, an inactive chiral analogue: implications for the mechanism of receptor binding.
  Biochemistry, 42, 12770-12783.
PDB codes: 1lw8 1pc1 1q4v
12817477 Z.Y.Guo, Y.H.Tang, S.Wang, and Y.M.Feng (2003).
Contribution of the absolutely conserved B8Gly to the foldability of insulin.
  Biol Chem, 384, 805-809.  
11742127 B.Xu, Q.X.Hua, S.H.Nakagawa, W.Jia, Y.C.Chu, P.G.Katsoyannis, and M.A.Weiss (2002).
A cavity-forming mutation in insulin induces segmental unfolding of a surrounding alpha-helix.
  Protein Sci, 11, 104-116.
PDB code: 1k3m
12196530 Q.X.Hua, Y.C.Chu, W.Jia, N.F.Phillips, R.Y.Wang, P.G.Katsoyannis, and M.A.Weiss (2002).
Mechanism of insulin chain combination. Asymmetric roles of A-chain alpha-helices in disulfide pairing.
  J Biol Chem, 277, 43443-43453.
PDB code: 1lkq
11923277 T.Kjeldsen, S.Ludvigsen, I.Diers, P.Balschmidt, A.R.Sorensen, and N.C.Kaarsholm (2002).
Engineering-enhanced protein secretory expression in yeast with application to insulin.
  J Biol Chem, 277, 18245-18248.  
11814349 Z.Y.Guo, L.Shen, and Y.M.Feng (2002).
The different folding behavior of insulin and insulin-like growth factor 1 is mainly controlled by their B-chain/domain.
  Biochemistry, 41, 1556-1567.  
12186542 Z.Y.Guo, L.Shen, and Y.M.Feng (2002).
The different energetic state of the intra A-chain/domain disulfide of insulin and insulin-like growth factor 1 is mainly controlled by their B-chain/domain.
  Biochemistry, 41, 10585-10592.  
11517220 M.A.Weiss, Q.X.Hua, W.Jia, S.H.Nakagawa, Y.C.Chu, S.Q.Hu, and P.G.Katsoyannis (2001).
Activities of monomeric insulin analogs at position A8 are uncorrelated with their thermodynamic stabilities.
  J Biol Chem, 276, 40018-40024.  
18763100 Y.Huang, Z.Liang, and Y.Feng (2001).
The relationship between the connecting peptide of recombined single chain insulin and its biological function.
  Sci China C Life Sci, 44, 593-600.  
11258877 Z.S.Qiao, Z.Y.Guo, and Y.M.Feng (2001).
Putative disulfide-forming pathway of porcine insulin precursor during its refolding in vitro.
  Biochemistry, 40, 2662-2668.  
11112528 M.A.Weiss, Q.X.Hua, W.Jia, Y.C.Chu, R.Y.Wang, and P.G.Katsoyannis (2000).
Hierarchical protein "un-design": insulin's intrachain disulfide bridge tethers a recognition alpha-helix.
  Biochemistry, 39, 15429-15440.  
11123908 S.H.Nakagawa, H.S.Tager, and D.F.Steiner (2000).
Mutational analysis of invariant valine B12 in insulin: implications for receptor binding.
  Biochemistry, 39, 15826-15835.  
  10091652 G.Kurapkat, M.Siedentop, H.G.Gattner, M.Hagelstein, D.Brandenburg, J.Grötzinger, and A.Wollmer (1999).
The solution structure of a superpotent B-chain-shortened single-replacement insulin analogue.
  Protein Sci, 8, 499-508.
PDB code: 1bzv
10397800 H.Berchtold, and R.Hilgenfeld (1999).
Binding of phenol to R6 insulin hexamers.
  Biopolymers, 51, 165-172.  
10497806 N.P.wing, and C.J.Cassady (1999).
Effects of cysteic acid groups on the gas-phase reactivity and dissociation of [M + 4H]4+ ions from insulin chain B.
  J Am Soc Mass Spectrom, 10, 928-940.  
  9631292 G.Dodson, and D.Steiner (1998).
The role of assembly in insulin's biosynthesis.
  Curr Opin Struct Biol, 8, 189-194.  
9708987 J.L.Whittingham, D.J.Edwards, A.A.Antson, J.M.Clarkson, and G.G.Dodson (1998).
Interactions of phenol and m-cresol in the insulin hexamer, and their effect on the association properties of B28 pro --> Asp insulin analogues.
  Biochemistry, 37, 11516-11523.
PDB codes: 1zeg 1zeh 1zei
9148904 C.Kristensen, T.Kjeldsen, F.C.Wiberg, L.Schäffer, M.Hach, S.Havelund, J.Bass, D.F.Steiner, and A.S.Andersen (1997).
Alanine scanning mutagenesis of insulin.
  J Biol Chem, 272, 12978-12983.  
9566117 C.McInnes, and B.D.Sykes (1997).
Growth factor receptors: structure, mechanism, and drug discovery.
  Biopolymers, 43, 339-366.  
8999905 D.C.Mynarcik, P.F.Williams, L.Schaffer, G.Q.Yu, and J.Whittaker (1997).
Analog binding properties of insulin receptor mutants. Identification of amino acids interacting with the COOH terminus of the B-chain of the insulin molecule.
  J Biol Chem, 272, 2077-2081.  
9228034 D.C.Mynarcik, P.F.Williams, L.Schaffer, G.Q.Yu, and J.Whittaker (1997).
Identification of common ligand binding determinants of the insulin and insulin-like growth factor 1 receptors. Insights into mechanisms of ligand binding.
  J Biol Chem, 272, 18650-18655.  
  9070440 G.Kurapkat, E.De Wolf, J.Grötzinger, and A.Wollmer (1997).
Inactive conformation of an insulin despite its wild-type sequence.
  Protein Sci, 6, 580-587.  
9131992 I.Pittman, S.H.Nakagawa, H.S.Tager, and D.F.Steiner (1997).
Maintenance of the B-chain beta-turn in [GlyB24] insulin mutants: a steady-state fluorescence anisotropy study.
  Biochemistry, 36, 3430-3437.  
9145374 J.Brange, L.Andersen, E.D.Laursen, G.Meyn, and E.Rasmussen (1997).
Toward understanding insulin fibrillation.
  J Pharm Sci, 86, 517-525.  
9245420 S.T.Huang, W.E.Choi, C.Bloom, M.Leuenberger, and M.F.Dunn (1997).
Carboxylate ions are strong allosteric ligands for the HisB10 sites of the R-state insulin hexamer.
  Biochemistry, 36, 9878-9888.  
9235985 X.Chang, A.M.Jorgensen, P.Bardrum, and J.J.Led (1997).
Solution structures of the R6 human insulin hexamer,.
  Biochemistry, 36, 9409-9422.
PDB codes: 1ai0 1aiy
  8931138 E.De Wolf, R.Gill, S.Geddes, J.Pitts, A.Wollmer, and J.Grötzinger (1996).
Solution structure of a mini IGF-1.
  Protein Sci, 5, 2193-2202.
PDB code: 1b9g
8844327 G.E.Sosinsky (1996).
Molecular organization of gap junction membrane channels.
  J Bioenerg Biomembr, 28, 297-309.  
7588759 D.R.Hodgson, F.E.May, and B.R.Westley (1995).
Mutations at positions 11 and 60 of insulin-like growth factor 1 reveal differences between its interactions with the type I insulin-like-growth-factor receptor and the insulin receptor.
  Eur J Biochem, 233, 299-309.  
  8011176 A.G.Schepky, A.M.Schmidt, T.Schmidt, P.Schulz-Knappe, and W.G.Forssmann (1994).
Determination of sulfated peptides by differential iodination.
  Biol Chem Hoppe Seyler, 375, 201-203.  
  7527339 H.Terasawa, D.Kohda, H.Hatanaka, K.Nagata, N.Higashihashi, H.Fujiwara, K.Sakano, and F.Inagaki (1994).
Solution structure of human insulin-like growth factor II; recognition sites for receptors and binding proteins.
  EMBO J, 13, 5590-5597.  
7809046 O.Gursky, E.Fontano, B.Bhyravbhatla, and D.L.Caspar (1994).
Stereospecific dihaloalkane binding in a pH-sensitive cavity in cubic insulin crystals.
  Proc Natl Acad Sci U S A, 91, 12388-12392.  
8136024 S.Q.Hu, G.T.Burke, and P.G.Katsoyannis (1993).
Contribution of the B16 and B26 tyrosine residues to the biological activity of insulin.
  J Protein Chem, 12, 741-747.  
1322139 J.Murray-Rust, A.N.McLeod, T.L.Blundell, and S.P.Wood (1992).
Structure and evolution of insulins: implications for receptor binding.
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1477273 O.Gursky, J.Badger, Y.Li, and D.L.Caspar (1992).
Conformational changes in cubic insulin crystals in the pH range 7-11.
  Biophys J, 63, 1210-1220.
PDB codes: 1aph 1bph 1cph 1dph
1961250 Q.X.Hua, S.E.Shoelson, M.Kochoyan, and M.A.Weiss (1991).
Receptor binding redefined by a structural switch in a mutant human insulin.
  Nature, 354, 238-241.
PDB codes: 1hit 1hiu
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time. Where a reference describes a PDB structure, the PDB codes are shown on the right.